Cat No.: 32380 ★Download Datasheet★ ★MSDS★
Introduction
Insulin is a peptide hormone exclusively produced in pancreatic beta-cells. It consists of A chain and B chain, which are linked by two disulfide bridges. Insulin is the primary hormone responsible for glucose metabolism. Impaired insulin secretion and insulin
resistance are key causes of type 2 diabetes (T2D).
Principle of the Assay
This assay is a two-site ELISA. The micro-plate is pre-coated with a monoclonal antibody against insulin. Standards and samples are added into the wells and coincubated with a monoclonal antibody conjugated to horseradish peroxidase (HRP) enzyme. After wash
step to remove any unbound substances, TMB substrate is added and colour develops in proportion to the amount of insulin bound
initially. The assay is stopped and the optical density of the wells determined using a micro-plate reader. Since the increases in
absorbance are directly proportional to the amount of captured insulin, the unknown sample concentration can be interpolated from a reference curve included in each assay.
Assay Performance
A.Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each assay.
Insulin (ng/mL) | Absorbance (450 nm) | Blanked Absorbance |
0 | 0.061 | 0 |
0.025 | 0.07 | 0.009 |
0.075 | 0.096 | 0.035 |
0.2 | 0.223 | 0.162 |
0.5 | 0.77 | 0.709 |
1.5 | 2.852 | 2.791 |
B.Sensitivity
The lowest insulin level that can be measured by this assay is 0.025 ng/mL.
C. Precision
Intra-assay Precision (Precision within an assay) C.V. < 2.9%.
Inter-assay Precision (Precision between assays) C.V.<4.9%.
D. Recovery
The recovery of the assay was determined by adding various amounts insulin to a sample. The measured concentration of the
spiked sample in the assay was compared to the expected concentration. The average recovery was 92%.
E. Specificity
Percent of cross reactivity
Human insulin: 100%
Rat insulin: 100%
Publications Citing This Product
1. Shu L, Hoo RL, Wu X, Pan Y, Lee IP, Cheong LY, Bornstein SR, Rong X, Guo J, Xu A. A-FABP mediates adaptive thermogenesis by promoting intracellular activation of thyroid hormones in brown adipocytes. Nature communications. 2017 Jan 27;8(1):1-6.
2. Fan W, Zhang R, Han D, Jiang Z, Li S, Zhang J, Li Y, Wang Y, Cao F. Reduced Sirtuin1 signalling exacerbates diabetic mice
hindlimb ischaemia injury and inhibits the protective effect of a liver X receptor agonist. Journal of cellular and molecular medicine.
2020 May;24(10):5476-90.
3. Wang B, Lin H, Li X, Lu W, Kim JB, Xu A, Cheng KK. The adaptor protein APPL2 controls glucose-stimulated insulin secretion via F-actin remodeling in pancreatic β-cells. Proceedings of the National Academy of Sciences. 2020 Nov 10;117(45):28307-15.
4. Zhang J, Ni Y, Qian L, Fang Q, Zheng T, Zhang M, Gao Q, Zhang Y, Ni J, Hou X, Bao Y. Decreased Abundance of Akkermansia
muciniphila Leads to the Impairment of Insulin Secretion and Glucose Homeostasis in Lean Type 2 Diabetes. Advanced Science.
2021 Jun 4:2100536.
5. So WY, Liu WN, Teo AK, Rutter GA, Han W. Paired box 6 programs essential exocytotic genes in the regulation of
glucose-stimulated insulin secretion and glucose homeostasis. Science Translational Medicine. 2021 Jun 30;13(600):eabb1038.
6. Liu D, Gu J, Shao W, Pang J, Qian X, Jin T. Comparison of metabolic beneficial effects of Liraglutide and Semaglutide in male
C57BL/6J mice. Canadian Journal of Diabetes. 2021 Sep 8.